reaction mass of sodium 1-amino-4-[[3,5-bis[[(chloroacetyl)amino]methyl]-2,4,6-trimethylphenyl]amino]-9,10-dihydro-9,10-dioxoanthracene-2-sulphonate and sodium 1-amino-4-[[3-[[(chloroacetyl)amino]methyl]-2,4,6-trimethylphenyl]amino]-9,10-dihydro-9,10-dioxoanthracene-2-sulphonate

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

other: data on Acid Blue 225_constituent 1

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Justification for type of information:

The experiment was conducted on one of the substance components. It should be noted that the lot tested was characterized by the Acid Blue 225_constituent 1 as main component; however, the impurity profile (which also includes the Acid Blue 225_constituent 2) resulted to be closely similar to that characterizing the substance under assessment (details are given in the document attached to IUCLID section 13.2).

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1974

Report date:

1974

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Principles of method if other than guideline:

The procedure adopted was the patch-test technique described in the "Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics" (1959) of the US Association of Food and Drug Officials (AFDO).

GLP compliance:

no

Test material

Test material information

Constituent 1

Reference substance name:

Acid Blue 225_constituent 1

IUPAC Name:

Acid Blue 225_constituent 1

Test animals

Species:

rabbit

Strain:

other: Russian Breed

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 1.5 to 2 kg.
- Housing: in order to prevent injuries by bites, the rabbits were kept separately in V2A wire cages.
- Diet: food (NAFAG, Gossau SG, rabbit food), ad libitum.
- Water: ad libitum.

Test system

Type of coverage:

occlusive

Preparation of test site:

other: intact and scarified

Vehicle:

other: 50 % polyethylene glycol trituration (PEG 400)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

0.5 g

Duration of treatment / exposure:

24 hours

Observation period:

72 hours

Number of animals:

3 males and 3 females

Details on study design:

TEST SITE
- Area of exposure: the shaven skin on the left side was slightly scarified immediately before treatment. A gauze patch 2.5 x 2.5 cm was soaked with the test substance and immediately applied to the prepared skin.
- Type of wrap if used: the patch was covered with a plastic film 5 x 5 cm, which was fixed to the body with adhesive tape.

OBSERVATION TIME POINTS
24 and 72 hours.

SCORING SYSTEM
The primary irritation index, as the measure of the acute skin irritation provoked by the substance, is defined as the average of all the ratings found after 24 and 72 hours on the intact and slightly scarified skin.

Erythema and Eschar Formation
No erythema 0
Very slight erythema (barely perceptible) 1
Well defined erythema 2
Moderate to severe erythema 3
Severe erythema (beet redness) to slight eschar formation (injuries in depth) 4

Oedema Formation
No oedema 0
Very slight oedema (barely perceptible) 1
Slight oedema (edges of area well defined by definite raising 2
Moderate oedema (raised approximately 1 mm) 3
Severe oedema (raised more than 1 mm and extending beyond area of exposure) 4

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

erythema score

Basis:

animal: 3/3

Remarks:

both intact and scarified skin

Time point:

24/48/72 h

Score:

< 2.3

Remarks on result:

no indication of irritation

Results 2

Irritation parameter:

edema score

Basis:

animal: 3/3

Remarks:

both intact and scarified skin

Time point:

24/48/72 h

Score:

< 2.3

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

The primary irritation index of test item was 0.

Any other information on results incl. tables

Evaluation of the Skin Reactions

Animal No. Sex	Reaction	24 hrs after application		72 hrs after application
		Intact skin	Scarified skin	Intact skin	Scarified skin
1 M	Erythema	0	0	0	0
	Oedema	0	0	0	0
2 M	Erythema	0	0	0	0
	Oedema	0	0	0	0
3 M	Erythema	0	0	0	0
	Oedema	0	0	0	0
4 F	Erythema	0	0	0	0
	Oedema	0	0	0	0
5 F	Erythema	0	0	0	0
	Oedema	0	0	0	0
6 F	Erythema	0	0	0	0
	Oedema	0	0	0	0

Applicant's summary and conclusion

Interpretation of results:

other: not classified, according to the CLP Regulation (EC) No 1272/2008

Conclusions:

Non irritating

Executive summary:

A study was performed to determine the skin irritation potential of test item on Russian breed rabbits according to "Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics" (1959) of the US Association of Food and Drug Officials (AFDO).

3 males and 3 females rabbits were shaved on the whole back and flanks two days before treatment with an electric clipper. The shaven skin on the left side was slightly scarified immediately before treatment. The test item was applied to each side in quantities of 0.5 g; before application a 50 % polyethylene glycol trituration (PEG 400) was made.

The primary irritation index as the measure of the acute skin irritation provoked by the substance is defined as the average of all the ratings found after 24 and 72 hours on the intact and slightly scarified skin.

The primary irritation index, as the measure of the acute irritation to the skin of rabbits, was found to be 0; therefore test item resulted to be as non-irritant to the skin of rabbits.

Discussion and conclusion

The scoring system criteria used to record the severity of the skin reactions followed in the current test are the same as those reported into the OECD guideline, therefore it is possible to re-evaluate the results in accordance with the CLP Regulation (EC) No 1272/2008.

None of the animals showed any observable response to treatment throughout the 72 hours observation period, in both the cases of intact and scarified skin. Therefore, it can be concluded that the test item does not meet the criteria to be classified as irritating, according to the CLP Regulation (EC) No 1272/2008.